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The peppermint breath test: a benchmarking protocol for breath sampling and analysis using GC-MS
Laura Di Francesco, Denise Biagini, Tommaso Lomonaco, Francesca G. Bellagambi, Sven Schuchardt, Olaf Holz, Katie Hamshere, Iain R White, Maxim Wilkinson, Stephen J. Fowler, 2020, original scientific article

Abstract: Exhaled breath contains hundreds of volatile organic compounds (VOCs) which offers the potential for diagnosing and monitoring a wide range of diseases. As the breath research field has grown, sampling and analytical practices have become highly varied between groups. Standardisation would allow meta-analyses of data from multiple studies and greater confidence in published results. The Peppermint Consortium has been formed to address this task of standardisation. In the current study we aimed to generate initial benchmark values for thermal desorption-gas chromatography-mass spectrometry (TD-GC-MS) analysis of breath samples containing peppermint-derived VOCs. Headspace analysis of peppermint oil capsules was performed to determine compounds of interest. Ten healthy participants were recruited by three groups. Each participant provided a baseline breath sample prior to taking a peppermint capsule, with further samples collected at 60, 90, 165, 285 and 360 min following ingestion. Sampling and analytical protocols were different for each institution, in line with their usual practice. Samples were analysed by TD-GC-MS and benchmarking values determined for the time taken for detected peppermint VOCs to return to baseline values. Sixteen compounds were identified in the capsule headspace. Additionally, 2,3-dehydro-1,8-cineole was uniquely found in the breath samples, with a washout profile that suggested it was a product of peppermint metabolism. Five compounds (α-pinene, β-pinene, eucalyptol, menthol and menthone) were quantified by all three groups. Differences in recovery were observed between the groups, particularly for menthone and menthol. The average time taken for VOCs to return to baseline was selected as the benchmark and were 441, 648, 1736, 643 and 375 min for α-pinene, β-pinene, eucalyptol, menthone and menthol respectively. An initial set of easy-to-measure benchmarking values for assessing the performance of TD-GC-MS systems for the analysis of VOCs in breath is presented. These values will be updated when more groups provide additional data.
Found in: ključnih besedah
Keywords: Volatile organic compounds, breath, diagnostics, standardisation
Published: 11.12.2020; Views: 100; Downloads: 0
.pdf Fulltext (1,10 MB)

Detection and quantification of exhaled volatile organic compounds in mechanically ventilated patients–comparison of two sampling methods
Iain R. White, Pouline M. van Oort, Waqar Ahmed, Craig Johnson, Jonathan Bannard-Smith, Timothy Felton, Lieuwe D. Bos, Royston Goodacre, Paul Dark, Stephen J. Fowler, 2020, original scientific article

Abstract: Exhaled breath analysis is a promising new diagnostic tool, but currently no standardised method for sampling is available in mechanically ventilated patients. We compared two breath sampling methods, first using an artificial ventilator circuit, then in “real life” in mechanically ventilated patients on the intensive care unit. In the laboratory circuit, a 24-component synthetic-breath volatile organic compound (VOC) mixture was injected into the system as air was sampled: (A) through a port on the exhalation limb of the circuit and (B) through a closed endo-bronchial suction catheter. Sorbent tubes were used to collect samples for analysis by thermal desorption-gas chromatography-mass spectrometry. Realistic mechanical ventilation rates and breath pressure–volume loops were established and method detection limits (MDLs) were calculated for all VOCs. Higher yields of VOCs were retrieved using the closed suction catheter; however, for several VOCs MDLs were compromised due to the background signal associated with plastic and rubber components in the catheters. Different brands of suction catheter were compared. Exhaled VOC data from 40 patient samples collected at two sites were then used to calculate the proportion of data analysed above the MDL. The relative performance of the two methods differed depending on the VOC under study and both methods showed sensitivity towards different exhaled VOCs. Furthermore, method performance differed depending on recruitment site, as the centres were equipped with different brands of respiratory equipment, an important consideration for the design of multicentre studies investigating exhaled VOCs in mechanically ventilated patients.
Found in: ključnih besedah
Summary of found: ...Exhaled breath analysis is a promising new diagnostic tool,...
Keywords: Volatile organic compounds, infection, breath, ventilator associated pneumonia
Published: 10.12.2020; Views: 104; Downloads: 0
.pdf Fulltext (1,61 MB)

Real-time multi-marker measurement of organic compounds in human breath: Towards fingerprinting breath
Iain R White, Kerry A Willis, Christopher Whyte, Rebecca Cordell, Robert S Blake, Andrew J Wardlaw, 2013, original scientific article

Abstract: The prospects for exploiting proton transfer reaction-time of flight-mass spectrometry (PTR-ToF-MS) in medical diagnostics are illustrated through a series of case studies. Measurements of acetone levels in the breath of 68 healthy people are presented along with a longitudinal study of a single person over a period of 1 month. The median acetone concentration across the population was 484 ppbV with a geometric standard deviation (GSD) of 1.6, whilst the average GSD during the single subject longtitudinal study was 1.5. An additional case study is presented which highlights the potential of PTR-ToF-MS in pharmacokinetic studies, based upon the analysis of online breath samples of a person following the consumption of ethanol. PTR-ToF-MS comes into its own when information across a wide mass range is required, particularly when such information must be gathered in a short time during a breathing cycle. To illustrate this property, multicomponent breath analysis in a small study of cystic fibrosis patients is detailed, which provides tentative evidence that online PTR-ToF-MS analysis of tidal breath can distinguish between active infection and non-infected patients.
Found in: ključnih besedah
Keywords: Volatile Organic Compounds, breath, proton transfer reaction mass spectrometry, Cystic Fibrosis
Published: 22.07.2019; Views: 807; Downloads: 0
.pdf Fulltext (1,37 MB)

Headspace volatile organic compounds from bacteria implicated in ventilator-associated pneumonia analysed by TD-GC/MS
Stephen J Fowler, Roy Goodacre, Iain R White, Tamara M E Nijsen, Waqar M Ahmed, Howbeer Muhamadali, Oluwasola Lawal, 2018, original scientific article

Abstract: Ventilator-associated pneumonia (VAP) is a healthcare-acquired infection arising from the invasion of the lower respiratory tract by opportunistic pathogens in ventilated patients. The current method of diagnosis requires the culture of an airway sample such as bronchoalveolar lavage, which is invasive to obtain and may take up to seven days to identify a causal pathogen, or indeed rule out infection. While awaiting results, patients are administered empirical antibiotics; risks of this approach include lack of effect on the causal pathogen, contribution to the development of antibiotic resistance and downstream effects such as increased length of intensive care stay, cost, morbidity and mortality. Specific biomarkers which could identify causal pathogens in a timely manner are needed as they would allow judicious use of the most appropriate antimicrobial therapy. Volatile organic compound (VOC) analysis in exhaled breath is proposed as an alternative due to its non-invasive nature and its potential to provide rapid diagnosis at the patient's bedside. VOCs in exhaled breath originate from exogenous, endogenous, as well as microbial sources. To identify potential markers, VAP-associated pathogens Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Staphylococcus aureus were cultured in both artificial sputum medium and nutrient broth, and their headspaces were sampled and analysed for VOCs. Previously reported volatile markers were identified in this study, including indole and 1-undecene, alongside compounds that are novel to this investigation, cyclopentanone and 1-hexanol. We further investigated media components (substrates) to identify those that are essential for indole and cyclopentanone production, with potential implications for understanding microbial metabolism in the lung.
Found in: ključnih besedah
Summary of found: ...Volatile organic compound (VOC) analysis in exhaled breath is proposed as an alternative due to...
Keywords: bacteria, exhaled breath, infection, ventilator-associated pneumonia, volatile organic compounds
Published: 18.07.2019; Views: 658; Downloads: 0

Breathomics and its Application for Disease Diagnosis: A Review of Analytical Techniques and Approaches
Enzo A Palombo, Konstantinos A Kouremenos, Ding Y Oh, Iain R White, David J Beale, Avinash V Karpe, Oliver A H Jones, 2018, independent scientific component part or a chapter in a monograph

Abstract: The application of metabolomics to an ever-greater variety of sample types is a key focus of systems biology research. Recently, there has been a strong focus on applying these approaches toward the rapid analysis of metabolites found in non-invasively acquired samples, such as exhaled breath (also known as ‘breathomics’). The sampling process involved in collecting exhaled breath is nonintrusive and comparatively low-cost. It uses a series of globally approved methods and provides researchers with easy access to the metabolites secreted by the human body. Owing to its accuracy and rapid nature, metabolomic analysis of breath is a rapidly growing field that has proven effective in detecting and diagnosing the early stages of numerous diseases and infections. This review discusses the various collection and analysis methods currently applied in breathomics research. Some of the salient research completed in this field to date is also assessed and discussed in order to provide a basis for possible future scientific directions.
Found in: ključnih besedah
Summary of found: non-invasively acquired samples, such as exhaled breath (also known as ‘breathomics’). The sampling process...
Keywords: Metabolomics, breath research, VOCs, breathomics
Published: 22.07.2019; Views: 816; Downloads: 0
.pdf Fulltext (25,96 MB)

Capturing and Storing Exhaled Breath for Offline Analysis
Stephen J Fowler, Iain R White, 2019, independent scientific component part or a chapter in a monograph

Abstract: In this chapter we will summarize and discuss methods for the capture and storage of exhaled breath, prior to offline (and indirect online) analysis. We will detail and compare methods currently in use, including their applications, key strengths, and limitations. In synthesizing the best features of each technique, we will propose an ideal standardized breath sampling solution, and give a personal vision on the next steps to be taken in this exciting area of breath research.
Found in: ključnih besedah
Keywords: Breath analysis, Breath sampling, Offline analysis, Thermal desorption, Gas chromatography-mass spectrometry
Published: 22.07.2019; Views: 814; Downloads: 0
.pdf Fulltext (36,24 MB)

Exhaled breath metabolomics reveals a pathogen-specific response in a rat pneumonia model for two human pathogenic bacteria: a proof-of-concept study
Iain R White, Pouline M van Oort, 2019, original scientific article

Abstract: Volatile organic compounds in breath can reflect host and pathogen metabolism and might be used to diagnose pneumonia. We hypothesized that rats with Streptococcus pneumoniae (SP) or Pseudomonas aeruginosa (PA) pneumonia can be discriminated from uninfected controls by thermal desorption-gas chromatography-mass-spectrometry (TD-GC-MS) and selected ion flow tube-mass spectrometry (SIFT-MS) of exhaled breath. Male adult rats (n = 50) received an intratracheal inoculation of 1) 200 µl saline, or 2) 1 × 107 colony-forming units of SP or 3) 1 × 107 CFU of PA. Twenty-four hours later the rats were anaesthetized, tracheotomized, and mechanically ventilated. Exhaled breath was analyzed via TD-GC-MS and SIFT-MS. Area under the receiver operating characteristic curves (AUROCCs) and correct classification rate (CCRs) were calculated after leave-one-out cross-validation of sparse partial least squares-discriminant analysis. Analysis of GC-MS data showed an AUROCC (95% confidence interval) of 0.85 (0.73-0.96) and CCR of 94.6% for infected versus noninfected animals, AUROCC of 0.98 (0.94-1) and CCR of 99.9% for SP versus PA, 0.92 (0.83-1.00), CCR of 98.1% for SP versus controls and 0.97 (0.92-1.00), and CCR of 99.9% for PA versus controls. For these comparisons the SIFT-MS data showed AUROCCs of 0.54, 0.89, 0.63, and 0.79, respectively. Exhaled breath analysis discriminated between respiratory infection and no infection but with even better accuracy between specific pathogens. Future clinical studies should not only focus on the presence of respiratory infection but also on the discrimination between specific pathogens.
Found in: ključnih besedah
Keywords: biomarkers, exhaled breath analysis, infection, pneumonia
Published: 22.07.2019; Views: 755; Downloads: 0
.pdf Fulltext (320,58 KB)

Circadian rhythm of exhaled biomarkers in health and asthma
Stephen Fowler, Royston Goodacre, David Ray, Dave Singh, Iain White, John Blaikley, Andrew Loudon, Robert Maidstone, Max Wilkinson, Hannah Durrington, 2019, original scientific article

Abstract: Circadian rhythms control many biological processes in the body in both health and disease. Greater understanding of diurnal variability in disease related biomarkers is crucial for their application in clinical practice and biomarkers of circadian rhythm are required to facilitate further research into disturbed chronicity. To determine if fractional exhaled nitric oxide and breath volatile biomarkers vary rhythmically during the day in healthy and asthmatic individuals. Ten individuals with moderate, atopic asthma (on regular inhaled corticosteroids) and 10 healthy volunteers (all non-smokers) completed an overnight visit where their exhaled breath volatiles and forced exhaled nitric oxide levels were collected every 6 h. Breath volatiles were analysed using gas chromatography mass spectrometry, after trapping these volatiles on sorbent materials for thermal desorption. Nine breath volatiles (including acetone and isoprene) exhibit diurnal variation across all individuals. Furthermore the circadian pattern of several VOCs is altered in individuals with asthma and fractional exhaled nitric oxide is rhythmic in asthma but not in healthy controls. Markers of circadian rhythm can be identified in breath and may offer insight into circadian profiling to help treat disease. Additionally this work suggests that time of day must be controlled when designing future biomarker discovery studies. Further work is required with larger cohorts to validate and extend these findings.
Found in: ključnih besedah
Summary of found: ...determine if fractional exhaled nitric oxide and breath volatile biomarkers vary rhythmically during the day...
Keywords: VOCs, breath, asthma, circadian
Published: 21.10.2019; Views: 775; Downloads: 0
.pdf Fulltext (479,14 KB)

Effects of high relative humidity and dry purging on VOCs obtained during breath sampling on common sorbent tubes
Maxim Wilkinson, Iain R White, Roy Goodacre, Tamara Nijsen, Stephen Fowler, 2020, original scientific article

Abstract: Offline breath analysis by thermal desorption gas chromatography mass spectrometry (TD-GC-MS) requires the use of sorbent traps to concentrate and store volatile compounds. The selection of which sorbent to use and best practices for managing high relative humidity are important considerations to allow for reproducible, untargeted, biomarker discovery in water saturated breath samples. This work aims to assess three commonly used sorbent materials for their use in breath volatile sampling and determine how the high relative humidity inherent in such samples effects the capture of volatile compounds of interest. TenaxGR, TenaxTA/Carbograph1TD and TenaxTA/Carbograph5TD tubes were selected as they are the most commonly used sorbents in the breath sampling literature. The recovery of 29 compounds in a standard mix loaded using high humidity gas was tested for each sorbent and compared to loading in dry gas. Water retention and dry purge rates were determined for each sorbent for 500 ml and 1000 ml breath collections. Finally, breath samples were collected simultaneously on to each sorbent type using the ReCIVA and analysed by TD-GC-MS. All three sorbents exhibited acceptable reproducibility when loaded with the standard mix in dry gas (RSD < 10%). Loading the standard mix in humid gas led to reduced recovery of compounds based on their chemical properties. Dry purging performance for each sorbent material was assessed and was shown to be 1.14, 1.13 and 0.89 mg H2O min−1 for TenaxGR, TenaxTA/Carbograph1TD and TenaxTA/Carbograph5TD respectively when flushed with 50 ml min−1 of N2. A comparison of breath profiles on different sorbents showed differences in background artefacts (sulfur dioxide, cyclopenten-1-one and 3-nonene) and endogenous breath compounds (2-methyl-furan and furfural). This work demonstrates that high relative humidity during sampling reduces the ability of sorbent tubes to capture volatile compounds and could impact method detection limits during breath sampling. Sufficient water to impair accurate analysis was retained on all tubes. Minimal differences were observed between sorbent materials when used to sample breath, however, suggestions are provided for sorbent selection for future studies.
Found in: ključnih besedah
Keywords: VOCs, Breath sampling, ReCIVA
Published: 27.07.2020; Views: 447; Downloads: 17
.pdf Fulltext (1,18 MB)

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